Component 2 Continuity of Life
All organisms are related through their evolutionary history
Spec Statement
the classification of organisms
into groups based on their
evolutionary relationships and
(a) that classification places
organisms into discrete and
hierarchical groups with other
closely related species
the need for classification and its
(b)
tentative nature
the three domain classification
system as compared with the
five Kingdom classification
system
(c)
(d) the characteristic features of
Kingdoms: Prokaryotae,
Protoctista, Plantae, Fungi,
Animalia
Comment
Candidates are required to understand the hierarchy of the taxons including Kingdom, Phylum, Class,
Order, Family, Genus, Species and be able to apply to given information.
Classification may change as additional information becomes available.
Recent biochemical evidence has shown that the kingdom prokaryotae should be split into two separate
groups based on some fundamental biochemical differences. All other organisms have eukaryotic cells.
This has led to the development of a scheme of classification which suggests all organisms evolved along
three distinct lineages, these are called domains. The organisms of each domain share a distinctive,
unique pattern of ribosomal RNA, which establishes their close evolutionary relationship. The three
domains of life are:
 Bacteria (or Eubacteria) which are the true bacteria.
 The Archaea (or Archaeabacteria) include the extremophile prokaryotes.
 Eukarya/ Eukaryota which include all eukaryotic organisms i.e. animals, plants, fungi and
protoctista.
Extremophiles exist in a wide variety of environmental conditions including extremes of temperature, pH,
salinity and pressure.
Prokaryotae - composed of prokaryotic cells, which lack a nuclear envelope and membrane-bound
organelles (the cell wall does not contain cellulose or chitin);
Plantae - multicellular eukaryotes, photosynthetic, cellulose cell wall;
Animalia - multicellular eukaryotes, no cell wall, heterotrophic; nervous coordination;
Fungi - heterotrophic eukaryotes, cell walls of chitin, most have filaments called hyphae; reproduce by
spores.

the use of physical features and
biochemical methods to assess
the relatedness of organisms,
including DNA ‘genetic
(e)
fingerprinting’ and enzyme
studies to show relatedness
without the problem of
morphological convergence
(f) the concept of species
the use of the binomial system in
(g)
naming organisms
biodiversity as the number and
(h) variety of organisms found within
a specified geographic region
biodiversity varying spatially and
(i) over time and affected by many
factors
biodiversity can be assessed in a
habitat e.g. Simpson’s Diversity
Index
(j)
biodiversity can be assessed
within a species at a genetic
level by looking at the variety of
(k) alleles in the gene pool of a
population, i.e. the proportion of
polymorphic loci across the
genome
biodiversity can be assessed at a
(l) molecular level using DNA
fingerprinting and sequencing
(m) biodiversity has been generated
Protoctista - mainly single cell eukaryotes, no tissue differentiation
Only the principle of genetic profiling is required here - biochemical methods measure the proportion of
DNA or proteins shared between species to estimate relatedness. DNA fragments or proteins are usually
displayed as bands on an electrophoresis gel. Biochemical methods can reduce the mistakes made in
classification due to convergent evolution.
Homologous features have evolved from the same original structure for different functions, e.g. the limbs
of reptiles, birds and mammals (pentadactyl limb), while analogous structures have evolved from different
structures to form the same function (wings of birds and insects).
Definition of biodiversity: the number of species and the number of individuals of each species in a given
environment.
Genetic, environmental and human factors affect biodiversity.
To investigate the biodiversity of a habitat, ecologists need to count the number of species present
(species richness) and the number of individuals within each species population.
It is possible to calculate the diversity of a habitat by using an index of diversity, such as Simpson’s
diversity index. Any value calculated using Simpson’s diversity index ranges between 0 and 1, the greater
∑ n(n−1)
the value, the greater the sample diversity. The formula to be used is D=1−
N (N −1)
Polymorphism is the word used to describe the presences of several different forms or types of individuals
among the members of a single species. Polymorphism results from the presence of polymorphic genes,
i.e. multiple alleles for the same gene.
Genetic biodiversity can be assessed by determining the:
 number of alleles at a locus e.g. T/t, IA/IB/IO
 proportion of the population that have a particular allele
Due to the difficulties involved in counting every single allele in a population researchers collect samples
of DNA and analyse the base sequences to look for variations between individuals. The greater the
variation in the base sequence, the greater the genetic diversity of the species.
Candidates need to appreciate the role of selective predation in natural selection but the detailed
 through natural selection
the different types of adaptations
of organisms to their
(n) environment including
anatomical, physiological and
behavioural adaptations
mechanism of evolution resulting in speciation is not required at AS level.
All species are uniquely adapted to the environment they inhabit; these adaptations include anatomical,
physiological and behavioural adaptations. For example animals that live in deserts have to cope with
extreme temperature fluctuations and a limited availability of water. Different organisms have different
adaptations to overcome the same problem.

SPECIFIED PRACTICAL WORK

 Investigation into biodiversity in a habitat

Genetic information is copied and passed onto to daughter cells

Spec Statement Comment

interphase and the main Candidates should be able to recognise the stages of mitotic cell division from diagrams and photographs.
(a) stages of mitosis They should be able to describe and explain the processes occurring at each stage.

the significance of mitosis as a Description of the process of mitosis in both plant and animal cells.
process in which daughter
(b) cells are provided with
identical copies of genes and
the process of cytokinesis
the significance of mitosis in No knowledge of oncogenes is required, but may be used as an application of unrestricted cell division
terms of damage and disease:
repeated cell renewal, damage
(c)
repair and healing and
unrestricted division leading to
cancerous growth
the main stages of meiosis Candidates should be able to recognise the meiotic stages from diagrams and photographs.
(names of subdivisions of They should be able to describe and explain the processes occurring at each stage.
(d)
prophase 1 not required) and
cytokinesis
the differences between This should include: number of nuclear divisions in the process, number of cells formed; ploidy of parental
mitosis and meiosis, including cells/nuclei; ploidy of daughter cells/nuclei; genetic nature of daughter cells/nuclei; pairing of homologous
(e) that mitosis produces non- chromosomes; crossing over; and segregation of homologous chromosomes.
identical daughter cells

SPECIFIED PRACTICAL WORK

 Scientific drawing of cells from slides of root tip to show stages of mitosis

 Scientific drawing of cells from prepared slides of developing anthers to show stages of meiosis
Sexual reproduction in humans

Spec Statement Comment

the structure and Candidates should be able to label and know the functions of parts of the male and female reproductive systems to
function of the include:
reproductive systems in Male:
(a) humans, including the
scrotum, testes, epididymis, vas deferens, seminal vesicle, prostate gland, urethra, penis;
examination of histology
of ovary and testis Female:
ovary, Fallopian tubes (oviducts), uterus, endometrium, cervix, vagina
(b) the processes of Spermatogenesis
spermatogenesis and  takes place in the seminiferous tubules
oogenesis to produce  candidates should know the types and stages of cell division which produce mature spermatozoa including:
spermatozoa and germinal epithelium; spermatogonia; primary and secondary spermatocytes; spermatids; spermatozoa and
secondary oocytes; be able to recognise these cells together with interstitial cells (cells of Leydig) and Sertoli cells on diagrams
sexual intercourse; and sections through a testis
fertilisation and  interstitial cells secrete testosterone which is involved in stimulating the process of spermatogenesis and
implantation
Sertoli cells are involved in providing nourishment and in protecting cells produced in this process
 candidates should be able to recognise and know the functions of the structures found in a mature sperm
cell.

Oogenesis
 oogenesis up to the secondary oocyte stage takes place in the ovary.
 candidates should know the types and stages of cell division which produce secondary oocytes including:
germinal epithelium; oogonia; primary and secondary oocytes; first polar body; primary, secondary and
Graafian follicles and corpus luteum. They should be able to recognise these structures on a diagram
showing stages in the development of a follicle from a primary follicle to a mature Graafian follicle, ovulation
and the subsequent development of the corpus luteum
 candidates should be able to recognise the structures found in and associated with a secondary oocyte
following ovulation to include: the corona radiata; zona pellucida; first polar body; cell membrane; cortical
granules and chromosomes / spindle apparatus suspended at metaphase II of meiosis.

Fertilization
  following sexual intercourse spermatozoa move into the Fallopian tubes
 capacitation increases the permeability of the membrane in front of the acrosome
 on contact with the zona pellucida the acrosome reaction releases hydrolase enzymes which digest the zona
pellucida
 the membranes of the sperm and secondary oocyte fuse and the genetic material of the sperm cell enters the
secondary oocyte
 this triggers the cortical reaction in which cortical granules fuse with the cell membrane and their contents
modify the zona pellucida to form the fertilisation membrane and prevents polyspermy
 entry of the genetic material also triggers meiosis II to continue forming the ovum and the second polar body
 the nuclei of the sperm and ovum fuse to form a zygotic nucleus

Implantation
• the zygote undergoes repeated mitotic divisions, called cleavage, to form a ball of cells called the blastocyst
• the blastocyst is moved into the uterus where it attaches and sinks into the endometrium – implantation
• a placenta forms between the tissues of the mother and the foetus.
(c) the endocrine control of Candidates should know and understand the role of hormones in reproduction in the female to include:
reproduction in the Menstrual Cycle:
female: including the • FSH secreted by the anterior pituitary gland stimulates the maturation of a follicle and stimulates the production
menstrual cycle, birth of oestrogen;
and lactation by • following menstruation the level of oestrogen, secreted by the developing follicle, increases in the blood which
reference to follicle triggers the repair of the endometrium; this inhibits FSH production and stimulates LH production;
stimulating hormone, • a high level of LH, secreted by the anterior pituitary, initiates ovulation; causes the Graafian follicle to develop
luteinising hormone, into a corpus luteum
oestrogen, • progesterone, secreted by the corpus luteum, causes further development of the endometrium prior to
progesterone, oxytocin menstruation
and prolactin and human • if implantation does not occur, falling FSH and LH levels cause the corpus luteum to degenerate, progesterone
chorionic gonadotrophin levels fall, the endometrium breaks down and is lost during menstruation; FSH secretion is no longer inhibited
the role of the placenta and another menstrual cycle is initiated.
including hormonal
control Candidates should be able to interpret a graphical representation of hormonal changes during the menstrual cycle in
relation to the development of the endometrium, ovulation and the role of negative feedback in controlling these
events.

Pregnancy:
• just before and following implantation, the developing embryo secretes HCG which maintains the corpus
luteum for the first 16 weeks of pregnancy
• the placenta then secretes progesterone and oestrogen which rise to high levels in the plasma
• FSH and LH secretion are inhibited
• progesterone suppresses the uterine wall’s ability to contract
(d) • oestrogen stimulates the growth of the uterus to accommodate the growing foetus and stimulates the growth and
development of the mammary glands during pregnancy;

Birth:
• just before birth oestrogen levels increase and progesterone levels decrease – the uterine wall can now
contract
• oxytocin secreted by the posterior pituitary gland stimulates contraction of the uterine wall which stimulates the
secretion of more oxytocin – this is an example of positive feedback
• prolactin is also released from the anterior lobe of the pituitary gland during and after birth to stimulate the
production of milk by the mammary glands

Candidates should be able to recognise and understand the roles of the different structures found in the placenta
including: counter-current flow between maternal and foetal blood supplies; chorionic villi; intervillous spaces;
transport to and from the foetus via the umbilical arteries and vein.

The role of the placenta in terms of: exchange of gases and nutrients; providing a barrier between the maternal and
foetal blood; protection from the immune system of the mother and from the difference in maternal and foetal blood
pressures; secretion of hormones.

The role of the amniotic fluid in acting as a shock absorber and in protecting the foetus during development.
Sexual reproduction in plants

Spec Statement
Comment
(a) the generalised structure Candidates should be able to recognise and label a half-flower of a typical regular, diocotyledonous, insect-
of flowers to be able to pollinated flower to include: receptacle, calyx, sepal, corolla, petal, stamen, filament, anther, carpel, ovary, ovule,
compare wind and style and stigma.
insect pollinated flowers They should be able to identify differences between an insect and a wind-pollinated flower in terms of function of
flower parts and adaptations to different methods of pollination.
(b) the development of The role of mitosis and meiosis in the development of pollen grains in an anther to include:
pollen and ovules, • mitosis to produce large numbers of pollen mother cells followed by meiosis to produce a tetrad of four haploid
including examination of cells;
prepared slides of anther • the role of the tapetum in pollen grain development;
and ovary • development and structure of a mature pollen grain, including subsequent mitotic divisions of the nucleus;
• dehiscence and pollen dispersal.

The role of mitosis and meiosis in the development of an ovule in the ovary to include:
• meiosis of a megaspore mother cell in the nucellus to produce four haploid megaspores;
• the growth and subsequent development of one of the megaspores including three mitotic divisions to produce
eight haploid nuclei within the embryo sac.

Candidates should be able to recognise structures in a mature ovule to include: funicle, integuments, micropyle,
embryo sac, female gamete, two synergids, two polar nuclei, three antipodal cells.

(c) cross and self-pollination Pollination is the transfer of pollen from an anther to a stigma. Candidates should understand the genetic
consequences of self-pollination and cross-pollination and appreciate how meiosis and random fertilisation result in
increased genetic variation through cross-pollination.
There are different adaptations of flowers that promote cross-pollination. These include irregular flower structure and
chemical self-incompatibility.
Candidates should understand the events following pollination to include:
• mitosis of the pollen grain nucleus to produce two male gametes and a pollen tube nucleus
• germination of a pollen grain on a compatible stigma;
• growth of a pollen tube (under the control of the pollen tube nucleus) through the digestion of the style through
the secretion of hydrolase enzymes
• entry of the pollen tube into the embryo sac through the micropyle
 Both male gametes are involved in separate fertilisation events:
(d) the process of double • one male gamete enters the embryo sac and fuses with the female gamete to produce a diploid zygote
fertilisation • the second male gamete fuses with the two polar nuclei to form a triploid primary endosperm nucleus

(e) the formation and The events that take place following double fertilisation to produce seeds and fruits include:
structure of seed and • the ovule developing into a seed;
fruit as shown by broad • the diploid zygote divides by mitosis to form the diploid embryo, consisting of plumule, radicle and one or two
bean and maize cotyledons;
• the triploid endosperm nucleus divides by mitosis to form endosperm tissue, an important food storage tissue in
cereal grains, e.g. wheat;
• the integuments develop into the testa;
• the micropyle remains as a pore in the testa
• the ovary wall develops into a fruit wall enclosing the seeds.
Candidates should be able to identify and label diagrams of broad bean and maize seeds to include: hilum (scar of
the funicle), micropyle, testa, position of radicle, plumule, cotyledons.
Seeds have evolved as a survival strategy for a terrestrial mode of life. Plants have developed different mechanisms
to enable the dispersal of seeds. This reduces competition following germination and increase the chance of growth
into mature plants.
(f) the process of Seeds can remain dormant until suitable conditions are present, i.e. availability of water, oxygen and a suitable
germination of Vicia faba temperature. Germination involves the rapid onset of biochemical activity and growth of a seedling until the plant can
(broad bean) carry out photosynthesis and become independent of the food stores contained in the cotyledons.
The stages of germination in a non-endospermic seed, e.g. broad bean include:
• water being imbibed through the micropyle; the cotyledons swelling and the testa being split to allow entry of
(g) the effect of gibberellin more oxygen for aerobic respiration;

• food reserves from the cotyledons, starch and proteins, are mobilised through hydrolysis (and also lipids in some
seeds);

• providing sources of energy for respiration and growth of the plumule and radical.
In endospermic seeds, e.g. maize, gibberellin, a plant hormone, is involved in the process of germination:
• following imbibition of water gibberellin is released by the embryo and diffuses to the aleurone layer which
contains proteins;

• gibberellins induces the production of hydrolytic enzymes, e.g. amylase which break down stored nutrients
• glucose and other nutrients diffuse to the embryo where they are used in aerobic respiration and growth.
Inheritance

Spec Statement Comment

(a) alleles as different forms Candidates should know, understand and use genetic terms to include: gene, locus, alleles, dominant, recessive,
of the same gene codominant, phenotype, genotype, homozygous, heterozygous, F1, F2, autosomes and sex chromosomes.

(b) the principles of Candidates should understand

monohybrid Mendelian • how Mendel used the results of experimental genetic crosses to derive his laws of inheritance and be able to
inheritance including apply these laws when solving genetic problems;
simple crosses involving
• the use of symbols to represent dominant, recessive, codominant and sex-linked alleles and how to represent
codominance
genetic crosses in diagrammatic form when solving genetics problems;
(c) the principles of dihybrid
Mendelian inheritance • how and why test crosses may be carried out;
including simple crosses • that Mendel’s laws only apply if genes are not linked, i.e. on different chromosomes and that if genes show
involving linkage linkage, the results of crosses will not follow the expected Mendelian ratios.
Candidates should be able to apply their knowledge and understanding of meiosis to explain the production of
recombinants through independent assortment of non-linked genes and how crossing over can produce
recombinants in linked genes.

(d) the use of a chi squared Candidates should understand under what conditions the Chi2 test can be used and that the Chi2 test can be used to
test determine if the results of a genetic cross are significantly different to expected results or whether the differences are
due to chance alone. They should know how to carry out and interpret the results of this test as follows:
• formulate a null hypothesis;
• calculate expected numbers from Mendelian ratios;
• calculate degrees of freedom;
• choose a suitable probability level;
• identify a Chi2 value from a Chi2 distribution table;
• accept or reject the null hypothesis.

(e) sex linkage as illustrated Candidates should understand sex-linkage in organisms with X and Y sex chromosomes, as the inheritance of a
by haemophilia and gene present on the X chromosome only. They should understand the significance of the lack of a corresponding
Duchenne muscular allele on the Y chromosome in terms of expression of recessive alleles.
dystrophy
(f) gene mutation as Mutations are spontaneous random events and that mutation rates are normally very low, but in organisms with short
illustrated by sickle cell life cycles and more frequent cell division, the rate of mutation is higher. It is the source of genetic variation which
anaemia and can result in evolution through natural selection. Most mutations occur during crossing over in prophase-I and non-
chromosome mutation disjunction in anaphase-I and anaphase-II.
as illustrated by Down's
syndrome
Mutations can affect protein synthesis and can change the phenotype of an organism, but some mutations have no
(g) the effect of mutagens,
effect on the phenotype. Gene (point) mutations affect single bases in a gene and chromosomal mutations affect
carcinogens and
many genes. The rate of mutation may be increased by mutagens, including ionising radiations (gamma radiation,
oncogenes
UV and X- rays), and certain chemicals, such as polycyclic hydrocarbons in cigarette smoke. A mutagen which
causes cancer is called a carcinogen. Some genes called proto-oncogenes can mutate to become oncogenes which
are involved causing uncontrolled cell division to form a cancer.
(h) the control of gene Epigenetics:
expression by factors • DNA can be modified post-replication
other than changes in
• this does not change the DNA base sequence but changes the ability of a gene to be transcribed during protein
the DNA sequence; the
synthesis
study of this is called
epigenetics • addition of methyl groups to bases prevents those bases being recognised and reduces the ability of that gene to
be expressed
• the histone proteins used to organise the DNA in a chromosome can also be modified – if the histone coils more
tightly this can prevent gene expression or if it coils more loosely can increase gene expression
Different epigenetic modifications can occur in cells of the same tissue and in different tissues resulting in different
expression of the same gene in different parts of the same organism.
Variation and evolution

Spec Statement Comment

(a) genetic and Candidates should be able to explain the difference between continuous and discontinuous variation in terms of:
environmental factors • number of genes controlling a particular phenotype
producing variation
• the effect of environmental factors
between individuals
(b) variation as continuous
and discontinuous;
heritable and non-
heritable
(c) the effect of inter- and Competition, environmental and human factors place selective pressures on the survival of different phenotypes and
intra-specific competition hence breeding success.
on breeding success
and survival
(d) the impact of selective
agencies (e.g. supply of
food, breeding sites,
climate, human impact)
on the survival of
organisms
(e) the concept of gene The gene pool is the total of all alleles for all of the genes in a population.
pool and genetic drift Selection pressures can change the allele frequencies of the alleles present at a particular gene locus in a population
(f) the effect of selection and that allele frequency can be expressed either as a proportion or a percentage of the total number of copies of all
changing the frequency alleles for that gene.
of alleles in a population
(g) the use of the Hardy- The Hardy-Weinberg principle states that the frequencies of dominant and recessive alleles and genotypes will
Weinberg principle and remain constant from one generation to the next, if certain conditions remain true. These conditions include:
equation • a large population (100+ individuals);
(h) the conditions under • no selection for or against any phenotype;
which the Hardy- • random mating throughout the population;
Weinberg principle • no mutations;
applies • the population is isolated, i.e. no immigration or emigration.

Candidates should know to apply the Hardy-Weinberg principle to estimate frequencies of dominant ore recessive
alleles or of different genotypes of a characteristic in a population using
p2 + 2pq + q2  = 1
where p = frequency of the dominant allele (A)
q = frequency of the recessive allele (a)
p+q = 1.0
The three terms of this binomial expansion indicate the frequencies of the three genotypes:
p2  = frequency of AA (homozygous dominant)
2pq  = frequency of Aa (heterozygous)
2
q  = frequency of aa (homozygous recessive)
(i) the concepts of isolation Evolution, in terms of speciation, will not take place if the conditions under which the Hardy-Weinberg principle
and speciation applies do not change. Speciation can occur due to:
(j) the separation of • genetic drift in isolated population
populations by • the founder effect of disproportionate allele frequencies in small populations
geographical,
behavioural, • natural selection
morphological, seasonal Isolation can be allopatric or sympatric and can be effected under these situations.
and other isolation Candidates should be able to apply their knowledge and understanding of meiosis to explain hybrid sterility e.g. in
mechanisms including the mule, and hybrid fertility e.g. in wheat.
hybrid sterility Selection pressures can affect the survival of different phenotypes in a population e.g. selective predation,
(k) Darwin's theory of camouflage, mimicry. Only individuals which survive to reproductive age can pass on selected alleles to their
evolution that existing offspring, thus changing the allele frequencies over time.
species have arisen
through modification of
ancestral species by
natural selection
Application of reproduction and genetics
Spec Statement
(a) the Human Genome
Project and its extension
to the 100K Genome
Project
(b) the ethical issues
surrounding the use of
this knowledge and its
application to the
screening of embryos for
genetic disorders e.g.
cystic fibrosis,
Huntington's disease,
thalassaemia
(c) how the genomes from Genome projects have also been completed for a number of other species including chimpanzees and other primates
other organisms have allowing scientists to look at evolutionary relationships and to conserve species in the future.
also been sequenced
including the mosquito,
Anopheles gambiae and
the Plasmodium parasite
that it transmits and that
better methods to
control malaria may be
developed as a result
Comment
The intended purpose of the Human Genome and 100K Projects is to improve knowledge and understanding of
genetic disorders and improve their diagnosis and treatment. The Human Genome Project used ‘Sanger
Sequencing’ which sequences relatively small sections of DNA at a time (usually <1,000 bps). This process took a
long time. New techniques e.g. Next Generation Sequencers (NGS) can sequence an entire genome in just a few
hours. NGS is enabling scientists to study variation within the human genome amongst 100 000 people in the U.K.
This is known as the 100K genome project.
Candidates should understand:
 the ethical issues in terms of ownership of genetic information, potential discrimination, social stigmatisation and
misuse of the data.
 the identification of allele sequences has enabled scientists to scan a patient’s DNA sample for mutated
sequences and also to compare the sequence of DNA bases in a patient’s gene to a normal version of the gene.
 that the screening of embryos has been performed to detect the presence of disorders such as cystic fibrosis,
Huntington’s disease and thalassaemia.
 that there are a number of concerns regarding the possibility of routine screening for adult onset disorders such
as Alzheimer’s disease and some cancers.
 that screening of embryos has led to concerns over choosing alleles to ensure specific characteristics
 that there are concerns that the risks of discrimination and social stigmatization that could outweigh the benefits
of testing
 the use of genetic screening and the value of genetic counselling
 the concerns regarding the ownership of genetic information and its misuse.
 Malaria is transmitted by the mosquito Anopheles gambiae. Rapid evolution of insecticide resistance in the
species is hampering attempts to eradicate the disease which is responsible for over a million deaths per year.
 The malarial parasite, Plasmodium sp. has also developed multi-drug resistance. Details of the life cycle of the
mosquito or the parasite are not required.
 Sequencing of the Anopheles gambiae genome is allowing scientists to develop chemicals, which could render
the mosquito susceptible again to insecticides.
 Sequencing of the Plasmodium sp. genome is allowing for the development of more effective drugs
(d) the use of PCR and Polymerase Chain Reaction allows, the quantity of DNA to amplified for analysis. Gel electrophoresis can then be
electrophoresis to used in the analysis of the DNA by producing a DNA profile.
produce a genetic These techniques can be used to identify the DNA of an individual:
fingerprint; the forensic
 An individual’s DNA profile is different from that of other individuals.
use of genetic
fingerprinting  Exons are regions of DNA that code for proteins. Between exons are regions of noncoding DNA called introns
which contain blocks of repeated nucleotides. It is the number of times that these blocks (Short Tandem Repeats
or STRs) are repeated that produces the variation in individuals.
 A number of STRs are used to build up a unique fingerprint in UK.
 D7S280 is an example of a STR where ‘GATA’ bases repeat on human chromosome 7. Different alleles of this
locus have from 6 to 15 tandem repeats of this sequence. The more times it repeats, the larger the fragment of
DNA will be.
 The polymerase chain reaction (PCR) is used to amplify small sections of DNA rapidly.
 PCR, is used to amplify the STRs by using a primer (single stranded DNA typically 6-25bp in length) which is
complimentary to the start of the sequence.
 PCR involves heating the DNA to 95oC to separate the two strands.
 The sample is then cooled to 50-60 oC to allow the primers to bind to the DNA strands (annealing).
 Heating to 70oC allows a thermally stable DNA polymerase (Taq) to add complimentary nucleotides (extension)
by forming the phosphodiester bonds in the sugar-phosphate backbone.
 This cycle is repeated. After 40 cycles over a billion copies of the target sequence can be produced from just one
piece of DNA.
 Gel electrophoresis is a method of separating DNA fragments according to size. The gel is made from agarose
(similar to agar), which contains pores in its matrix.
 DNA samples are loaded into wells at one end and a voltage is applied across the gel. DNA is attracted to the
positive electrode due to its negative charge on the phosphate group. Smaller fragments find it easier to migrate
through the pores in the gel and so travel further than large fragments in the same time.
 Fragment size can be estimated by using running a DNA ladder (which contains fragments of known size)
alongside.
Candidates should appreciate the limitations of these techniques and the ethical issues raised in their use.
(e) the formation of The processes by which recombinant plasmids can be engineered include:
recombinant DNA by • the role of restriction endonuclease enzymes and DNA ligase;
insertion of foreign DNA
• the significance of sticky ends;
into bacterial plasmids
and the cloning of the • the use of antibiotic resistance genes in the selection of recombinant bacteria.
bacteria to produce
useful molecules as Candidates should be able to describe how a fragment of DNA containing a human gene can be prepared:
illustrated by insulin • by using restriction endonuclease enzymes to cut out the gene from a human chromosome
or
• by extracting mRNA from a cell actively synthesising the required protein / polypeptide and using reverse
transcriptase and DNA polymerase to produce a double stranded cDNA fragment

Candidates should understand that producing cDNA overcomes the following problems:
• locating the gene;
• restriction enzymes cutting the gene into non-functional fragments;
• the presence of introns;
• the need for post-transcriptional processing to produce functional mRNA.

Candidates should be aware of the reasons why there are concerns over the genetic engineering of bacteria to
include:
• the use of antibiotic resistance genes in plasmids and the ready exchange of genetic material between bacteria;
• the possible transfer of antibiotic resistance genes to pathogenic bacteria;
• the possible transfer / activation of oncogenes by using fragments of human DNA.
(f) issues surrounding the Candidates should be able to evaluate the possible benefits of GM crop production against the concerns associated
use of gene technologies with the use of this technology including:
to produce genetically Benefits:
modified crops by
• superior keeping qualities;
inserting a gene from one
organism into another to • higher yield;
convey disease • a substantial reduction in pesticide use on crops engineered for resistance to fungal pathogens and insect
resistance e.g. in GM attack.
tomatoes or a desired Concerns:
characteristic e.g. in GM
 soya • dispersal of pollen from crops engineered for herbicide resistance to wild relatives;
• unknown effects of eating new protein produced in crop;
• a reduction in biodiversity.
(g) the advantages and
disadvantages of using Gene therapy can be used to treat genetic disorders by inserting functional DNA sequences into cells to counteract
gene therapy for the the effect of a defective gene.
treatment of disease as
 Genetic disease can be treated by replacing genes or replicating the function of genes using drugs.  
illustrated by muscular
dystrophy  There are two possible methods of replacing defective genes: somatic cell therapy and germ line therapy.
Somatic cell therapy will not prevent the condition being passed on and germ line therapy is very rare.
 The aim of gene therapy is to treat a genetic disease by replacing defective alleles in a patient with copies of a
new DNA sequence.
 Duchenne Muscular Dystrophy (DMD) is a recessive, sex linked form of Muscular Dystrophy affecting up to one
in 3 500 live male births.
 It is caused by a mutation in the dystrophin gene resulting in the failure to produce dystrophin, which is an
important structural component of muscle tissue. The result is severe wasting of the muscles and sufferers are
often wheelchair bound by the time they reach teenage years. Life expectancy is only 27.

 A drug called drisapersen has been developed which aims to treat DMD by introducing a 'molecular patch' over
the exon with the mutation making the gene readable again. A shorter from of dystrophin is produced, but one
thought to be more functional than the untreated version. This type of treatment is called exon skipping. 

(h) the use of genomics and Genomics is the study of the structure, function, evolution and mapping of genomes as exemplified by the Human
its possible impact on Genome and 100K Projects. This should enable healthcare to be improved by more accurate diagnosis, better
healthcare of the future prediction of the effect of drugs and improved design of drugs; new and improved treatments for disease. With the
introduction of NGS technology it may be possible to look at tailoring therapies to individual patients where an
individual could have a unique treatment for a common disease.

(i) the issues surrounding Candidates should be able to explain the term tissue engineering including the role of stem cells.
the use of stem cells for The advantages and disadvantages of using stem cells include:
replacing damaged
Advantages:
tissues and organs
• speed of production;
• large scale production;
• production of genetically identical cells / organisms.
Disadvantages:
• in mammals the technique is very expensive and unreliable;
• in plants disease / entry of pathogens may cause problems;
• inadvertent selection of disadvantageous alleles;
• long term / unforeseen effects such as premature aging.
There are ethical issues associated with obtaining stem cells from embryos and the cloning of human tissues and
organs.